Use of enhanced green fluorescent protein to optimize and quantitate infection of target cells with recombinant retroviruses.
نویسندگان
چکیده
Recombinant retroviral vectors are useful tools for gene transfer in both gene therapy and research applications. An enhanced form of green fluorescent protein has been incorporated into recombinant retroviruses as a marker to follow infected cells. In this paper, we extended the use of the fluorescent reporter to quantify protein expression using such analytical tools as fluorescent microscopy, flow cytometry and fluorescent plate reader analysis. These tools enabled us to rapidly assess the titer of recombinant retrovirus harvested from packaging cells and to optimize parameters for infection of different cell lines.
منابع مشابه
Influence of E1-Deleted Recombinant Adenoviruses on B7.1 and IL-2 Expression in C1498 Cells
Knowing that adenoviral vectors could initiate innate immunity, the ability of E1-deleted recombinant adenovirus (Ad-E1Δ) in induction of B7.1 and IL-2 molecules was studied. Methods: The expression of green fluorescent protein in C1498 cells following transfection of these cells with adenovirus green fluorescent protein vector confirmed the ability of adenovirus vectors in infecting the cells ...
متن کاملIn vitro Labeling of Neural Stem Cells with Poly-L-Lysine Coated Super Paramagnetic Nanoparticles for Green Fluorescent Protein Transfection
Background: The magnetic nanoparticle-based transfection method is a relatively new technique for delivery of functional genes to target tissues. We aimed to evaluate the transfection efficiency of rat neural stem cell (NSC) using poly-L-lysine hydrobromide (PLL)-coated super paramagnetic iron oxide nanoparticles (SPION). Methods: The SPION was prepared and coated with PLL as transfection agent...
متن کاملConstruction of Various γ34.5 Deleted Fluorescent-Expressing Oncolytic herpes Simplex type 1 (oHSV) for Generation and Isolation of HSV-Based Vectors
Background: Oncolytic herpes simplex virus (oHSV)-based vectors lacking γ34.5 gene, are considered as ideal templates to construct efficient vectors for (targeted) cancer gene therapy. Herein, we reported the construction of three single/dually-flourescence labeled and γ34.5-deleted, recombinant HSV-1 vectors for rapid generation and easy selection/isolation of different HSV-Based v...
متن کاملTransient expression of green fluorescent protein in radish (Raphanus sativus) using a turnip mosaic virus based vector
It is possible to use transgenic plants, as bioreactors, for the production of recombinant inexpensive chemicals and drug components. Transient gene expression is an appropriate alternative to stable transformation because it allows an inexpensive and rapid method for expression of recombinant proteins in plant tissues. In recent years, plant viral vectors have been increasingly developed as su...
متن کاملA Model to Study the Phenotypic Changes of Insect Cell Transfection by Copepod Super Green Fluorescent Protein (cop-GFP) in Baculovirus Expression System
Background: Baculovirus expression system is one of the most attractive and powerful eukaryotic expression systems for the production of recombinant proteins. The presence of a biomarker is required to monitor transfection efficiency or protein expression levels in insect cells. Methods: The aim of this study was to construct a baculovirus expression vector encoding a copepod super green fluore...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- BioTechniques
دوره 26 5 شماره
صفحات -
تاریخ انتشار 1999